Process For The Production Of An Enriched Natural Antioxidant Mixture From A Single Source Plant

ABSTRACT

A process for extracting antioxidants from a plant, including contacting a plant material from a guayusa plant for a first time with a solvent, thereby obtaining a first slurry, filtering said first slurry, thereby obtaining a first extract, contacting said plant material for a second time with said solvent, thereby obtaining a second slurry, filtering said second slurry, thereby obtaining a second extract, combining said first extract and said second extract, thereby generating an third extract containing at least antioxidants, xanthines, and amino acids, and substantially drying said third extract.

STATEMENT OF PRIORITY

The present application claims priority to U.S. Provisional ApplicationNo. 61/898,362, titled “Process For The Production Of An EnrichedNatural Antioxidant Mixture From A Single Source Plant” and filed Oct.31, 2013.

TECHNICAL FIELD

The present disclosure relates to the field of therapeutic naturalproducts and a process for producing an enriched extraction of a naturalantioxidant mixture rich in cinnamic acids, catechins, amino acids andxanthines from a single source plant material, guayusa, which can beused to prevent a host of inflammatory diseases such as diabetes,cancer, heart disease, Alzheimer's and obesity as well as help treatailments associated with poor glucose metabolism, endothelialdysfunction, oxidative stress, inflammation and cognitive decline.

BACKGROUND

Ilex guayusa is an Amazonian tree of the holly genus, native to theEcuadorian Amazon Rainforest. The plant yields xanthenes such ascaffeine. In addition to caffeine, guayusa also contains theobromine, astimulant commonly found in chocolate and L-theanine, a glutamic acidanalog found in green tee that has been shown to reduce physical andmental stress. See Kimura K, Ozeki M, Jeneja L, Ohira H (2007).“L-Theanine reduces psychological and physiological stress responses.”Biol Psychol 74 (1): 39-45. doi: 10.1016/j.biopsycho.2006.06.006 (http://dxdoi.org/10.1016%2Fj.biopsycho.2006.06.006). PMID16930802(//hwww.ncbi.nlm.nih. gov/pubmed/16930802).

Current approaches for the use of the guayusa plant include steeping theleaves and forming a beverage substrate (the “tea”) as previouslydisclosed in http://www.stashtea.com/info/guayusa.aspx andhttp://www.runa.org/our-guayusa/. The finished beverage produced fromboth the tea leaves and the Ready to Drink (“RTD”) beverages isdescribed as a naturally caffeinated herbal infusion produced from theleaves of a holly tree. The finished beverage composition containsantioxidants, catechins, vitamins and amino acids at relatively lowlevels (Antioxidant and Compounds Analysis of Guayusa tea, “Lab Number:056939”. Advanced Botanical Consulting & Testing, Inc., 2010) as well asxanthines or caffeine as a natural sources of energy, FIG. 1. See ORACAnalysis, Test Report for the Whole ORAC Values (Report). Genox. 2010,FIG. 2.

Accordingly, there is a need to find natural remedies for inflammatorydiseases and enhance cognitive function through the production of anextract which contains a specific ratio of actives that allow foreffective nutritional formulation and dosage in a concentrated way toensure product efficacy without having to consume large volumes ofliquid.

In addition, there is a need for an extract or essence of the guayusatea leaves with an enhanced finished product sensory profile and a shelflife extension. Specifically, a need exists for a masking agentstabilizing oxidative damage to the antioxidant content withoutbitterness in a finished tea based beverages (IFT 2013, Chicago, Ill.Presented by TEAWOLF).

Finally, there is need for a medicinal extract with the specific activeratio which provides for health benefits not yet seen or described byingestion of the guayusa tea alone. See Antioxidant and CompoundsAnalysis of Guayusa tea, “Lab Number: 056939”. Advanced BotanicalConsulting & Testing, Inc., 2010. Suggestive of their role in diseaseprevention, the active compounds in the guayusa plant, once harvestedand concentrated to the correct ratio as described within thedisclosure, demonstrate and provide enhanced beneficial effects in humanhealth.

SUMMARY OF THE INVENTION

According to one aspect, the present disclosure relates to a process forextracting antioxidants from a plant, including contacting a plantmaterial from a guayusa plant for a first time with a solvent, therebyobtaining a first slurry, filtering said first slurry, thereby obtaininga first extract, contacting said plant material for a second time withsaid solvent, thereby obtaining a second slurry, filtering said secondslurry, thereby obtaining a second extract, combining said first extractand said second extract, thereby generating an third extract containingat least antioxidants, xanthines, and amino acids, and substantiallydrying said third extract.

In another aspect, the present disclosure relates to an antioxidantmixture prepared by a process comprising the steps of: contacting aplant material from a guayusa plant for a first time with a solvent,thereby obtaining a first slurry, filtering said first slurry, therebyobtaining a first extract, contacting said plant material for a secondtime with said solvent, thereby obtaining a second slurry, filteringsaid second slurry, thereby obtaining a second extract, combining saidfirst extract and said second extract, thereby generating an thirdextract containing at least antioxidants, xanthines, and amino acids,and substantially drying said third extract.

BRIEF DESCRIPTION OF THE DRAWINGS

The following figures are included to illustrate certain aspects of thepresent invention, and should not be viewed as an exclusive embodiments.The subject matter disclosed is capable of considerable modification,alteration, and equivalents in form and function, as will occur to onehaving ordinary skill in the art and the benefit of this disclosure.

FIG. 1 is a prior art lab result, Antioxidant and Compounds Analysis ofGuayusa tea, “Lab Number: 056939”. Advanced Botanical Consulting &Testing, Inc., 2010.

FIG. 2 is another prior art lab result, ORAC Analysis, Test Report forthe Whole ORAC Values Report, Genox, 2010.

DETAILED DESCRIPTION Definitions

The term “filtration” is used to refer to either ultrafiltration ornanofiltration. In general, the “permeate” is the component that passesfreely through the filter, while the “retentate” is the component thatis retained by the filter.

The term “microfiltration” refers to processes that use filtrationmembranes having larger pore size than both ultrafiltration andnanofiltration. Microfiltration involves subjecting the antioxidantextract or eluate to filtration through a filter having a pore size ofless than about 0.50 μM.

As used herein, the term “nanofiltration” refers to processes that usefiltration membranes having a smaller molecular weight or pore size thanthose typically used in ultrafiltration processes. Like ultrafiltration,nanofiltration rejects a portion of the extract or eluate componentsabove a certain molecular size while allowing those of a smaller size topass through. Suitable nanofiltration membranes for use in the processdisclosed herein are preferably made from polymers having a nominalmolecular weight cut off of from about 700 Da to about 5000 Da(corresponding to pore sizes in the range of from about 17 A to about 40A). Particularly preferred nanofiltration membranes are made frompolymers having a nominal molecular weight cut off of from about 800 Dato about 2000 Da (corresponding to pore sizes in the range of from about18 A to about 27 A). This pore size, in general, allows unoxidizedphenolic compounds to pass through the membrane while retaining largersized protein like compounds.

As used herein, the term “plant material” means species of guayusa (Ilexguayusa), preferably the tea leaves.

As used herein, the terms “antioxidant-containing eluate” and“antioxidant eluate” are used interchangeably to refer to the desiredantioxidant-containing component collected after exposure to theadsorbent as described herein. Such adsorption step may be columnadsorption, or any other adsorption means known to those having ordinaryskill in the art.

As used herein, the term “nutrient-rich extract” means an extract havinga nutrient concentration of at least about 5 times, and preferably atleast about 15 times, the starting composition based on theHigh-Performance Liquid Chromatography (“HPLC”) measurement, of thedesired compounds (chlorogenic acids, xanthines, amino acids andflavonoids).

As used herein, the term “antioxidant containing retentate” means theamino acid-containing components remaining on the upstream side of thefilter after ultrafiltration or nanofiltration. Such aminoacid-containing retentates are used as the source of polyphenols,flavonoids, xanthines and other water soluble vitamins.

As used herein, the terms “amino acid-containing eluate,” “amino acideluate” and “amino acid rich extract” are used interchangeably to referto the desired amino acid-containing components collected after exposureto the adsorbent as described herein such as the L-theanine or luecine.Such adsorption step may be column adsorption, or any other adsorptionmeans known to those having ordinary skill in the art.

As used herein, the term “ultrafiltration” means a filtration methodthat uses an open filtration membrane with a pore size capable ofallowing through molecules from at least about 10,000 Da to at leastabout 100,000 Da in molecular weight. Typically, ultrafiltration removeslarge molecular weight polysaccharides and proteins, but not oxidizedphenolics.

As used herein, the term “water,” means any of: deionized water, reverseosmosis water, distilled water, process water, ion exchange or mixturesthereof.

All amounts, parts, ratios and percentages used herein are by weightunless otherwise specified.

Process

The first step in one embodiment of the presently disclosed processdisclosed herein involves blanching the leaves by contacting the plantmaterial with boiling water, preferably greater than 80° C., forapproximately 30 seconds. In some embodiments, this step may alsoutilize other leaf pretreatment methods such as steam blanching, citricacid blanching or a convention withering step. However, in preferableembodiments, optimum conditions are met with non withered leaves toavoid any un-necessary oxidation of the phytochemicals prior toextraction. The blanched leaves are then contacted with a solvent toobtain an extract comprising the soluble antioxidants (flavonoids andcinnamic acids), xanthines, and amino acids. The plant material used inthe present disclosure is guayusa (Ilex guayusa), and more preferablythe guayusa leaves (other Ilex species with the particular nutrientsdesired may also be used, i.e. Ilex vomitoria). The preferred extractionprocess involves steeping the guayusa leaves in and ethanol:hot watersolution. The guayusa leaves are extracted using a 70:30 ethanol:hotwater solution from about 70° C. to about 100° C. (from about 158 F. toabout 212° F.), preferably from about 75° C. to about 90° C. (from about167° F. to about 194° F.), and more preferably from about 80° C. toabout 85° C. (from about 176° F. to about 185° F.), and a combined tealeaf to solvent ratio of about from 1:10 to about 1:20, i.e. for every 1kg of guayusa leaves used, about 10 to 20 kg of solvent solution isused. Additionally, the extraction can be completed using a completewater extraction, supercritical extraction or similar however the yieldsof polyphenols and catechins and other active compounds will change.

The guayusa leaves are soaked in the solvent between approximately 60minutes to approximately 240 minutes, after which the wet leaves arefiltered out through one or more layers of cheese cloth, or othersimilar straining material, and the antioxidant extract is collected.The wet leaves may then be re-extracted numerous additional times(preferably two or three more times) with another volume of hot water ora water solvent mixture and soaked for about 60 minutes to about 120minutes further. The leaves are again filtered out and the nutrient richextract collected. The filtered nutrient extracts can then combined andare ready for further processing or the second nutrient rich extract canbe further processes itself, then combined with the first for a finishedextract with ideal product specifications.

Exposing Extract to Absorbent

In one embodiment, the nutrient rich extract resulting from the previousstep is subsequently exposed to an adsorbent, which separates thedesired compounds from other associated substances, such as the pectinsand fiber. The result is a nutrient rich containing eluate that issubstantially free of the aforementioned associated insoluble compoundsand impurities. The preferred method of carrying out this adsorptionstep is column chromatography. However, any similar method of separationcommonly known to those skilled in the art is acceptable. For example,the nutrient rich extract and adsorbent may be combined in a solventmedium and mixed thoroughly or through a multistage supercriticalextraction process.

As previously mentioned, column chromatography is the preferred methodto separate the desired extract from the other components in thenutrient rich extract. To separate via column chromatography, an inertcolumn, preferably one made of glass or plastic, is first packed with anadsorbent or column packing The adsorbent material may be any of avariety of hydrophobic cationic materials, however, polymeric resins,such as polyamides or polyclar are preferred. The column is thenequilibrated with a solvent that is preferably water-soluble and doesnot form two phases when mixed with water. The solvent utilized in thisphase of the process is preferably selected from water, ethanol,propylene glycol, glycerin, weak solutions of acetone, propanols, otherlike alcohols, and mixtures thereof. More preferably, the solventcomprises a mixture of water and ethanol. Still more preferably, thesolvent mixture comprises less than about 80%, preferably less thanabout 70% ethanol, by weight of the solvent. In an alternate embodiment,the solvent comprises water.

Next, the nutrient rich extract is pumped through the column and thecomponents that are not adsorbed, or poorly adsorbed, i.e. amino acids,will be the first class of components to elute with the solvent. As thesolvent strength is increased, such as, for example, through theaddition of more ethanol, more strongly adsorbed components are releasedfrom the adsorbent material in the column and elute with the solvent.This process allows for the separation of the desired materials and theproduction of an amino acid rich-containing eluate having a compositioncontaining at least approximately 5% total amino acids.

Filtration of the Amino Acid -Containing Eluate

The amino acid containing eluate is then subjected to a filtration step,to remove additional high molecular weight material, such aspolysaccharides, pectins and fiber, and further enrich the nutritionalconcentration of the eluate. As defined above, this filtration step maybe either ultrafiltration or nanofiltration. Each of these filtrationprocesses is set forth below

Nanofiltration involves contacting the amino acid eluate with ananofiltration membrane to provide a filtered nutrient-rich extract.Nanofiltration according to the present disclosure removes the highermolecular weight materials such as polysaccharides, pectins and fibers.

It is preferred that the nanofiltration step be carried out while thenutrient eluate is at a temperature of from about 30° C. to about 50° C.(about 86° F. to about 122° F.), preferably from about 35° C. to about50° C. (about 95° F. to about 122° F.), and more preferably from about45° C. to about 50° C. (about 113° F. to about 122° F.).

Efficient nanofiltration is typically achieved by warming the nutrienteluate after exposure to the adsorbent material and just prior tonanofiltration. The pressure at which nanofiltration is carried out ispreferably sufficiently high to provide adequate flow of the nutrienteluate through the membrane to achieve the desired processing. However,the pressure is preferably not so high as to remove substantial amountsof water from the system. According to the present disclosure,nanofiltration is typically carried out under a hydrostatic pressure offrom about 100 psi to about 300 psi, preferably from about 180 psi toabout 280 psi, applied to the upstream side of the membrane.

Suitable nanofiltration membranes for use in the process of the presentdisclosure are made from polymers having a nominal molecular weight cutoff of from about 700 Da to about 5000 Da (corresponding to pore sizesin the range of from about 17 A to about 40 A). Preferred nanofiltrationmembranes are made from polymers having a nominal molecular weight cutoff of from about 800 Da to about 2000 Da (corresponding to pore sizesin the range of from about 18 A to about 27 A).

Suitable polymers are those that have less affinity for the desiredpolyphenol I flavonoid components in the nutrient eluate. Polymers suchas cellulose and the like are usually suitable for making thesenanofiltration membranes.

Typically, the resulting amino acid-rich extract is cooled to atemperature of about 16° C. (about 60° F.) or less.

Similar to nanofiltration, ultrafiltration involves contacting thenutrient eluate with an ultrafiltration membrane to provide a filterednutrient-rich extract. Ultrafiltration uses an open filtration membranewith a pore size capable of allowing through molecules from at leastabout 10,000 Da to at least about 100,000 Da in molecular weight.

When utilizing ultrafiltration, the nutrient eluate can generally befiltered at a temperature of from about 30° C. to about 50° C. (about86° F. to about 122° F.), preferably from about 35° C. to about 50° C.(about 95° F. to about 122° F.), and more preferably from about 45° C.to about 50° C. (about 113° F. to about 122° F.).

Once the nutrient eluate is subjected to one of the aforementionedfiltration processes the nutrient composition obtained contains not lessthan approximately 10% total amino acids. The resulting amino acid-richextract can now be enriched with the antioxidant polyphenols andxanthines removed during the initial extraction and separation steps bycombining the two concentrated extracts, where evaporation and dryingcan produce a finished extract that contains no less than 30%chlorogenic acids, 10% xanthines and 5% amino acids, or 45 w/w % totalnutrients. More specifically, the finished extract contains no less than40% chlorogenic acids, 15% xanthines and 10% amino acids, or 65 w/w %total nutrients.

Using the Nutrient-Rich Extract

After subjecting the plant material to one of the foregoing embodiments,the resulting nutrient-rich extract may then be used to lower theglycemic response of a subject in a more effective way than priorguayusa compositional function (see Swanston-Flatt, S K et. Al Glycaemiceffects of traditional European plant treatments for diabetes. Studiesin normal and streptozotocin diabetic mice. Diabetes Res. 1989 February;10(2):69-73) and assist in mood enhancement, provide a natural source ofenergy, decrease the risk of cardiovascular disease, enhance brainfunction, assist with weight management and lower oxidative stress.

The specific composition produced can provide an optimum source ofnatural energy that contains both an active element for enhancedmetabolic function with its caffeine content while also assisting withglucose metabolism and regulation with the high level of chlorogenicacids. This ratio provides an ideal synergistic level of chlorogenicacids and caffeine to achieve effective weight loss benefits, whilststill shunting overactive insulin activity and hypertension typicallyassociated with an increase in caffeine consumption.

Further, the specific composition may provide a masking agent to controlthe astringent, brackish or bitter taste when formulating, add a levelof sweetness to the product or allow for increased shelf stability byprotecting or stabilizing the finished formulation from oxidation.

EXAMPLES Example 1

The following examples are illustrative of embodiments disclosed herein.Parts and percentages are by dry weight unless otherwise indicated. Itshould be noted that these examples describe a wide range of conditions,which together with the above descriptions, illustrate the presentdisclosure in a non limiting fashion.

About 400 g of commercial quality non withered guayusa tea leaves areextracted with about 1000 L of an ethanol:deionized water solution(70:30), at about 75° C. to about 85° C. (about 167° F. to about 186°F.) for approximately 120 minutes. The resulting slurry is filteredthrough two layers of muslin cloth and yields about 1200 g of crude teaextract. The residual leaves are re-extracted with deionized water underthe foregoing conditions and again filtered through muslin cloth threemore times. The tea extracts are combined and subjected to vacuum dryingto concentrate the extract and remove additional impurities at about 80°C. until less than 3% moisture. The above semi solid extract isdissolved in ethanol then the top layer decanted. The top layer is thendried in a rotary drum dryer until less than 2% moisture. The finalextract contains a profile of chlorogenic acids, catechins and caffeinefor use as a nutritional ingredient in food and beverage applications.

The final extract phytochemical profile is as seen in Table 1:

TABLE 1 Serial No. Phyto-constituent Assay in w/w % Analysis method 1Total Polyphenols 35 ± 0.5 w/w % Folin-Ciocalteu by spectrophotometricmethod 2 Total Chlorogenic 32 ± 0.35 w/w % LCMS/MS method acids 3Caffeine 26 ± 0.25 w/w % LCMS/MS method 4 Theobromine 0.0215 ± 0.005 w/w% LCMS/MS method 5 L-Theanine 0.011 ± 0.003 w/w % LCMS/MS method

Example 2

About 200 g of commercial quality guayusa tea leaves are extracted withabout 550 L of an ethanol:deionized water solution (70:30), at about 75°C. to about 85° C. (about 167° F. to about 186° F.) for approximately120 minutes. The resulting slurry is filtered through two layers ofmuslin cloth and yields about 1200 g of crude tea extract. The residualleaves are re- extracted with deionized water under the foregoingconditions and again filtered through muslin cloth. The tea extracts arecombined and subjected to microfiltration via a 0.45 μM pleated filter(1.5 ft.sup.2, acrylic co-polymer on a polypropylene-polyester support).The resulting permeate is then subjected to nanofiltration using aMillipore® 1000 Dalton Molecular Weight Cut Off (MWCO) filter. Theresulting permeate is then used to produce a clarified tea extract whichcontain the chlorogenic acids, xanthines and other phenols. Theretentates from these filtrations, which contain the amino acids andglycosides, are combined to yield an amino acid-containing retentatewith a volume of about 300 ml and about S0 Bx. This extract has anactive concentration of about 600 mg/L. The nutrient rich extract isdiluted with deionized water in a 1:1 ratio, and used as feed in thenext step.

Amberlite XAD 16HP® (Rohm & Haas) is packed in a column (2.5 cmID.times.75 cm height) to give a column volume of about 350 ml. Thecolumn is washed with about 4-5 column volumes of deionized water. Thediluted nutrient extract from the foregoing extraction step is pumpedinto the column until breakthrough occurs (about 100 ml). The column isfirst eluted with deionized water to remove the non-phenolic materialsincluding amino acids and other polysaccharides. When the eluate givesno precipitate or a clouding reaction with ethanol, the water elution isstopped. This eluate contains about 200 mg/L amino acids and about 10mg/ml tea polysaccharides. It is then nanofiltered using a 1000 DaltonMWCO membrane to separate the high molecular weight non-phenolicmaterials from amino acids. The amino acid-rich extract is then added tothe clarified polyphenol tea extract and further concentrated undervacuum until dry to remove any residual solvents and impurities. Thefinished substrate is then solubilized in a distilled solution and driedin a rotary evaporator to produce an extract of less than approximately3% moisture.

The final extract phytochemical profile is as seen in Table 2:

TABLE 2 Serial No. Phyto-constituent Assay in w/w % Analysis method 1Total Polyphenols 47.1 ± 1.5 w/w % Folin-Ciocalteu by spectrophotometricmethod 2 Total Chlorogenic 41.5 ± 0.35 w/w % LCMS/MS method acids 3Caffeine 36.8 ± 0.25 w/w % LCMS/MS method 4 Catechins 7.2 ± 0.05 w/w %LCMS/MS method 5 L-Theanine 1.40 ± 0.04 w/w % LCMS/MS method 6Theobromine 0.266 ± 0.007 w/w % LCMS/MS method

Example 3

200 grams of non-withered Guayusa tea leaves are extracted with anEthanol:water mixture (95:5) at 55° C. to about 65° C. The water to tealeaves ratio is about 15:1. This extraction is continued for about 2hours, and the resulting nutrient dense extract is filtered using afiltration funnel with a waterman filtering paper. The residual tealeaves are then extracted twice more with an ethanol:water solvent at aratio of 90:10, and this second and third nutrient extracts 2 and 3 arepassed through filtration using whatman filter paper to remove anyresidual tea powder residue. The three nutrient extracts are combined,and the total active content in them is determined to be about 220 mg/L.

The combined extracts are send through a buchi rotary evaporator vacuumpump to concentrate the extract and remove any residual solvents. Thesemi-solid extract is completely dried using a savant speedvac vacuumsystem. The final extract provides a nutrient dense substrate containingthe optimum combination of antioxidants and caffeine for use as anutritional component in foods, beverages and supplements.

The final extract phytochemical profile is as seen in Table 3:

TABLE 3 Serial No. Analysis Amount Method 1 Total Polyphenols 22.3 ± 1.5w/w % Folin-Ciocalteu by spectrophotometric method 2 Total Chlorogenic21.1 ± 0.35 w/w % LCMS/MS method acids 3 Caffeine 17.6 ± 0.15 w/w %LCMS/MS method 4 Catechins 1.5 ± 0.05 w/w % LCMS/MS method 5 L-Theanine0.002 ± 0.04 w/w % LCMS/MS method 6 Theobromine 0.165 ± 0.007 w/w %LCMS/MS method

Example 4

Seven female subjects, ages 29-45, were given 300 mg of guayusa extractcontaining 42% chlorogenic acid, 16% xanthines and 12% amino acidsorally for 3 weeks. Subjects were measured at the onset and upon finaladministration for Nuerotransmitter levels of Serotonin, GABA, Dopamine,Norepinephrine, Epinephrine, Glutamate and Creatinine Resultsdemonstrate an increase in Serotonin and GABA levels by 22% and 26%respectively, and an increase in of Dopamine, Norephipephrine,Epinephrine by 12%, 11% and 15%. Glutamate and Creatinine levelsremained unchanged.

Example 5

Two sets of six ready-to-drink tea beverages were prepared by steepingconventional green tea leaves in water at a 1:1 ratio for ten minutes,filtering, pasteurizing, then bottling. With one set, prior to bottling,200 mg of guayusa extract was added that contained 32% chlorogenic acid,16% xanthines and 5% amino acids and 30% total glycosides. Packaging wascarried out in conventional 500 ml RTD glass bottles and sealed withPTFE screw lined caps. The tea beverages were stored for a period of 12weeks in a temperature controlled light deprived container. One samplefrom each set was analyzed over the 12 weeks period for total flavonolglycoside concentration and total EGCG concentration.

Over the 12 weeks period, the control set containing no guayusa reducedtotal glycoside concentration by 37% and total EGCG concentration by 40%with most of the degration during the last two weeks of the study, week10-week 12. Over the same 12 week period the six green tea beveragescontaining the 200 mg of guayusa extract per 500 ml seen an averagedecrease in total glycosides of 2% and a total average decrease in EGCGof 8%. In addition sensory scores completed on both sets indicated anincrease in overall product taste as sweetness score by 92% of thesensory evaluations.

Although the present disclosure has been described with reference tospecific embodiments, this description is not meant to be construed in alimited sense. Various modifications of the disclosed embodiments, aswell as alternative embodiments of the disclosure will become apparentto persons skilled in the art upon the reference to the description ofthe present disclosure. It is, therefore, contemplated that the appendedclaims will cover modifications that fall within the scope of thedisclosure.

What is claimed is:
 1. A process for extracting antioxidants from aplant, comprising: contacting a plant material from a guayusa plant fora first time with a solvent, thereby obtaining a first slurry; filteringsaid first slurry, thereby obtaining a first extract; contacting saidplant material for a second time with said solvent, thereby obtaining asecond slurry; filtering said second slurry, thereby obtaining a secondextract; combining said first extract and said second extract, therebygenerating an third extract containing at least antioxidants, xanthines,and amino acids; and substantially drying said third extract.
 2. Theprocess of claim 1, wherein said plant material is non-withered guayusaleaves, thereby preventing loss of anti-oxidants.
 3. The process ofclaim 1, wherein said plant material is guayusa leaves that are one ofblanched or raw guayusa leaves.
 4. The process of claim 1, furthercomprising contacting at least one of said first or second extract withan absorbent prior to combining said first and second extracts, therebygenerating an amino acid containing eluate.
 5. The process of claim 4,further comprising filtering the amino acid containing eluate via one ofan ultrafiltration or nanofiltration.
 6. The process of claim 1, whereinsaid plant material is ground up portions of said guayusa plant leaves.7. The process of claim 1, wherein said solvent is an ethanol:deionizedwater solvent comprised of approximately a 70/30 ratio.
 8. The processof claim 1, wherein the solvent is selected from the group consisting ofwater, ethanol, and mixtures thereof.
 9. The process of claim 1, whereinsaid filtering is performed via a muslin cloth.
 10. The process of claim1, further comprising filtering said second extract for a second timeprior to combining said second extract with said first extract.
 11. Anantioxidant mixture prepared by a process comprising the steps of:contacting a plant material from a guayusa plant for a first time with asolvent, thereby obtaining a first slurry; filtering said first slurry,thereby obtaining a first extract; contacting said plant material for asecond time with said solvent, thereby obtaining a second slurry;filtering said second slurry, thereby obtaining a second extract;combining said first extract and said second extract, thereby generatingan third extract containing at least antioxidants, xanthines, and aminoacids; and substantially drying said third extract.
 12. The antioxidantmixture of claim 11, comprising a composition of no less than 30%chlorogenic acids, 10% xanthines, and 5% amino acids.
 13. Theantioxidant mixture of claim 11, comprising a composition of no lessthan 40% chlorogenic acids, 15% xanthines, and 10% amino acids.
 14. Theantioxidant mixture of claim 11, wherein said plant material isnon-withered guayusa leaves, thereby preventing loss of anti-oxidants15. The antioxidant mixture of claim 11, wherein said plant material isguayusa leaves that are one of blanched or raw guayusa leaves.
 16. Theantioxidant mixture of claim 11, wherein said process further comprisescontacting at least one of said first or second extract with anabsorbent prior to combining said first and second extracts, therebygenerating an amino acid containing eluate.
 17. The antioxidant mixtureof claim 11, wherein said process further comprises filtering the aminoacid containing eluate via one of an ultrafiltration or nanofiltration.18. The antioxidant mixture of claim 11, wherein said solvent is anethanol:deionized water solvent comprised of approximately a 70/30ratio.
 19. The antioxidant mixture of claim 11, wherein the solvent isselected from the group consisting of water, ethanol, and mixturesthereof.
 20. The antioxidant mixture of claim 11, wherein the processfurther comprises filtering said second extract for a second time priorto combining said second extract with said first extract.